Role of Kaposi's Sarcoma-Associated Herpesvirus C-Terminal LANA Chromosome Binding in Episome Persistence

Kelley-Clarke, Brenna; Leon-Vazquez, Erika De; Slain, Katherine; Barbera, Andrew J.; Kaye, Kenneth M.

doi:10.1128/jvi.02395-08

Cited by 35 publications

(42 citation statements)

References 86 publications

(138 reference statements)

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“…In agreement with previous results, LANA associated with mitotic chromosomes (yellow, resulting from the colocalization of green and red), with concentrations near telomeres and centromeres ( Fig. 2b and g; arrows indicate peritelomeric staining, and arrowheads indicate pericentromeric staining) (28,37). LANA⌬465-497 ( Fig.…”

Section: Resultssupporting

confidence: 92%

Identification of Kaposi's Sarcoma-Associated Herpesvirus LANA Regions Important for Episome Segregation, Replication, and Persistence

Vázquez

Carey

Kaye

2013

J Virol

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Kaposi's sarcoma-associated herpesvirus (KSHV) latency-associated nuclear antigen (LANA) is a 1,162-amino-acid protein that mediates the maintenance of episomal viral genomes in latently infected cells. The two central components of episome persistence are DNA replication with each cell division and the segregation of DNA to progeny nuclei. LANA self-associates to bind KSHV terminal-repeat (TR) DNA and to mediate its replication. LANA also simultaneously binds to TR DNA and mitotic chromosomes to mediate the segregation of episomes to daughter nuclei. The N-terminal region of LANA binds histones H2A and H2B to attach to mitotic chromosomes, while the C-terminal region binds TR DNA and also associates with chromosomes. Both the N-and C-terminal regions of LANA are essential for episome persistence. We recently showed that deletion of all internal LANA sequences results in highly deficient episome maintenance. Here we assess independent internal LANA regions for effects on episome persistence. We generated a panel of LANA mutants that included deletions in the large internal repeat region and in the unique internal sequence. All mutants contained the essential N-and C-terminal regions, and as expected, all maintained the ability to associate with mitotic chromosomes in a wild-type fashion and to bind TR DNA, as assessed by electrophoretic mobility shift assays (EMSA). Deletion of the internal regions did not reduce the half-life of LANA. Notably, deletions within either the repeat elements or the unique sequence resulted in deficiencies in DNA replication. However, only the unique internal sequence exerted effects on the ability of LANA to retain green fluorescent protein (GFP) expression from TR-containing episomes deficient in DNA replication, consistent with a role in episome segregation; this region did not independently associate with mitotic chromosomes. All mutants were deficient in episome persistence, and the deficiencies ranged from minor to severe. Mutants deficient in DNA replication that contained deletions within the unique internal sequence had the most-severe deficits. These data suggest that internal LANA regions exert critical roles in LANA-mediated DNA replication, segregation, and episome persistence, likely through interactions with key host cell factors.

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Section: Resultssupporting

confidence: 92%

Identification of Kaposi's Sarcoma-Associated Herpesvirus LANA Regions Important for Episome Segregation, Replication, and Persistence

Vázquez

Carey

Kaye

2013

J Virol

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“…Previous studies showed an interaction of LANA with histones, and more specifically the C terminus of LANA with the TR (8,(38)(39)(40)(41)(42)(43)(44). This prompted us to explore the role of H2AX (or ␥H2AX) in mediating the interaction between LANA and the TR.…”

Section: Expression Of Ddr Genes and Phosphorylation Status Of H2ax Dmentioning

confidence: 99%

H2AX Phosphorylation Is Important for LANA-Mediated Kaposi's Sarcoma-Associated Herpesvirus Episome Persistence

Jha

Upadhyay

et al. 2013

J Virol

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The DNA damage response (DDR) of host cells is utilized by a number of viruses to establish and propagate their genomes in the infected cells. We examined the expression of the DDR genes during Kaposi's sarcoma-associated herpesvirus (KSHV) infection of human peripheral blood mononuclear cells (PBMCs). The genes were mostly downregulated, except H2AX, which was upregulated during infection. H2AX is important for gammaherpesvirus infectivity, and its phosphorylation at serine 139 is crucial for maintenance of latency during mouse gamma-herpesvirus 68 (MHV-68) infection. We now also observed phosphorylation of H2AX at serine 139 during KSHV infection. H2AX is a histone H2A isoform shown to interact with the latency-associated nuclear antigen (LANA) encoded by KSHV. Here, we show that LANA directly interacted with H2AX through domains at both its N and C termini. The phosphorylated form of H2AX (␥H2AX) was shown to colocalize with LANA. Chromatin immunoprecipitation (ChIP) assays showed that a reduction in H2AX levels resulted in reduced binding of LANA with KSHV terminal repeats (TRs). Binding preferences of H2AX and ␥H2AX along the KSHV episome were examined by whole-episome ChIP analysis. We showed that ␥H2AX had a higher relative binding activity along the TR regions than that of the long unique region (LUR), which highlighted the importance

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“…Alternatively, although perhaps less likely, the apparently LANA -portion of the proliferating l B cells could be infected but be at an early stage prior to well-organized LANA tethering of the episome(s) to the chromatin. The MIFC algorithm queried punctate LANA dots and thus would not detect cells with LANA expressed at a low level or broadly distributed (46).…”

Section: Kshv-driven Growth/proliferation In Tonsillar B Cellsmentioning

confidence: 99%

KSHV infects a subset of human tonsillar B cells, driving proliferation and plasmablast differentiation

Hassman

Ellison²,

Kedes³

2011

J. Clin. Invest.

102

113

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Role of Kaposi's Sarcoma-Associated Herpesvirus C-Terminal LANA Chromosome Binding in Episome Persistence

Cited by 35 publications

References 86 publications

Identification of Kaposi's Sarcoma-Associated Herpesvirus LANA Regions Important for Episome Segregation, Replication, and Persistence

Identification of Kaposi's Sarcoma-Associated Herpesvirus LANA Regions Important for Episome Segregation, Replication, and Persistence

H2AX Phosphorylation Is Important for LANA-Mediated Kaposi's Sarcoma-Associated Herpesvirus Episome Persistence

KSHV infects a subset of human tonsillar B cells, driving proliferation and plasmablast differentiation

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